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. Author manuscript; available in PMC: 2013 Jul 12.
Published in final edited form as: Immunogenetics. 2007 Sep 21;59(10):813–821. doi: 10.1007/s00251-007-0250-6

Fig. 2.

Fig. 2

Nitr9L associates specifically with Dap12. a Predicted protein structures of recombinant zebrafish proteins utilized in cotransfection assays. Myc-tagged Nitr9-long (Nitr9L) is shown on the left, and FLAG-tagged Dap10 and Dap12 are shown on the right. Nitr9L encodes two extracellular Ig domains of the variable (V) and intermediate (I) types (Yoder et al. 2004). The cytoplasmic ITAM of Dap12 and the putative Dap10 signaling motif, YMNV, are indicated (Yoder et al. 2007). b AD-293 cells were cotransfected with plasmids encoding Myc-tagged Nitr9L and FLAG-tagged Dap10 or Dap12 as indicated (lanes 3–9). Cells were then preincubated with an anti-Myc antibody for cross-linking (X-link) before immunoprecipitation. pcDNA3 (lane 1) was used as a vector control, and isotype-matched antibody (lane 2) was used as a negative control for cross-linking. Immunoprecipitates (anti-Myc) were separated by SDS-PAGE and subjected to Western analyses using an anti-FLAG antibody (upper panel). Whole-cell lysates from the same treatments also were separated by SDS-PAGE and Western analyses completed with an anti-FLAG antibody to show the expression level of Dap10 and Dap12 (bottom panel)